Hi , recently i tried to knock out my gene of interest by CRISPR Cas9 KO Plasmid Santa cruz biotechnology kit in MDA-MB-231 cell line. After transfection i found that transfection efficiency is very low in this cells. Has anyone performed transfection to this cell line by this kit? Can anyone offer suggestions?
Maybe this new product:
Single-Cell Analysis Solution Single Cellome™ | Yokogawa Deutschland GmbH
could help. This system component automates the penetration and injection of single cells using a nanopipette. It‘s low invasiveness enables manipulation of live single cells. The system integrates with multiple manufacturers’ inverted microscopes. With a high success rate of approx. 95% success rate of injection.
thanks for your valuable information Erika . But we don't have this equipment in our lab.
Hi Mehdi Rabiee Valashedi ,
I did created a MDA-MB-231 KO cell line using this kit. I transfected it using Lipofectamine 3000 but still got low KO efficiency after one round of transfection. Then I transfected the same cells twice (2 rounds of transfections with the plasmids) before puromycine and FACS selection. I also did a clonal selection to finally get KO clones. It took a long time but I could get it. Hope it helps!
Thank you Manon Ros . How much puromycin did you use for cell selection?
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